Transcript Nutrition, Toxins and Health: Facts and Speculation Stephanie Seneff Wise Traditions Workshop Weston Price Foundation Monday, Nov.

Nutrition, Toxins and Health:
Facts and Speculation
Stephanie Seneff
Wise Traditions Workshop
Weston Price Foundation
Monday, Nov. 12, 2012
Part II: Biology
Download the slides from
http://people.csail.mit.edu/seneff/
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
Introduction
Modern Diseases
cancer
Alzheimer’s
cardiovascular disease
asthma
multiple sclerosis
depression
allergies
fibromyalgia
diabetes
ADHD
Parkinson’s disease
obesity
GERD
heart failure
autism
seizures
Why We are Sick
We need to worry about sulfur!
Current agricultural practices are depleting sulfate
from the soil
This leads to insufficient dietary sulfur, especially
for vegetarians
Environmental toxins deplete sulfur
Sulfate depletion in the body has
consequences
widespread
A Biological Definition of Aging
Excess glycation
damage to
proteins (AGEs)
Cholesterol and
sulfate depletion in
cell membranes
CELLS
Dysfunctional blood
system leading to blood
clots and hemorrhages
Increased susceptibility
to infection and cancer
Diabetes and Obesity
Dysfunction
Consequences
Cholesterol Transport
Sulfate Deficiency
Obesity
eNOS
SiNiC
Blood Dysfunction
Diabetes
Heart Disease
Cancer
Impaired Gut Bacteria
Infections
Impaired Autophagy
Dysfunction
Cholesterol Transport
Under healthy conditions, abundant cholesterol
sulfate is synthesized in the skin following sun
exposure, and this results in generous supply of
cholesterol and sulfate to the tissues.
Inadequate sun exposure and sunscreen
this process.
impair
Sulfate Deficiency
The body depends upon heparan sulfate
proteoglycans (HSPGs) as a
temporary
storage bin for glucose
Impaired sulfate synthesis in the skin disrupts this
process and leads to diabetes
Obesity
Insufficient HSPGs impairs muscles’ ability
utilize glucose as a fuel
to
Fat cells insinuate themselves into the loop by
transforming glucose to fat for later release to
supply safe usable fuel to muscles
eNOS
eNOS (endothelial Nitric Oxide Synthase) is a
“moonlighting” enzyme: it makes sulfate upon
sunlight stimulation, and switches to
nitric
oxide (nitrate) under stress
SiNiC
The slide from Sulfur into Nitrogen into Carbon as
we age
Oxygen transport in the blood depends upon these
three atomic elements (S, N, C)
which can
react with oxygen and form anions that stabilize
the blood and safely carry oxygen
Sulfur is the healthiest choice;
nitrogen is
associated with many pathologies; carbon acting
alone will lead to
acidosis and cancer
Consequences
Blood Dysfunction
The stability of the blood colloidal system depends
upon adequate cholesterol sulfate.
When sulfate is depleted, suspended cells and
particles become deficient in negative charge, and
start to stick together.
This leads to blood clots and hemorrhaging
Heart Disease
Cardiovascular plaque develops as an alternative
mechanism to produce cholesterol sulfate from
damaged LDL and homocysteine
Arthritis
Arthritis, and other chronic diseases like
Alzheimer’s and multiple sclerosis, result because
the body robs an organ system of sulfate in order
to keep the blood from coagulating
Cancer
Cancer develops as a mechanism to massively
convert glucose to lactate,
which is a much
safer fuel – especially when cells are insulin
resistant
Infection
Susceptibility to infection is a consequence of
impaired cholesterol sulfate synthesis, which
introduces widespread pathology
Infection serves a useful role in
resupplying
critical nutrients such as
cobalamin,
folate, and heparan sulfate
Impaired Autophagy
Autophagy is the process by which cells dispose of
the garbage that accumulates with living – they
can recycle misfolded proteins into new proteins
and replace broken mitochondria with fresh ones.
Excess nitric oxide leads to impaired autophagy,
which results in accumulation of debris and busted
mitochondria. Over time, the cell becomes so
impaired that it has to shut down.
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
Cholesterol Transport
Under healthy conditions, abundant cholesterol
sulfate is synthesized in the skin following sun
exposure, and this results in generous supply of
cholesterol and sulfate to the tissues.
Inadequate sun exposure and sunscreen
this process.
impair
Cholesterol is Essential for Mobility
and a Nervous System
• Plants contain no cholesterol
• Plants can’t move
• Plants don’t have a
nervous system
• In a sense, cholesterol is to animals
as chlorophyll is to plants
A Provocative Proposal
• Cholesterol sulfate supplies
oxygen, sulfur, cholesterol,
energy
energy and negative charge to
the tissues
• Sulfate is synthesized from sulfide in skin and
blood stream utilizing energy in sunlight
– Protects from UV damage and keeps microbes out
• Endothelial Nitric Oxide Synthase
(eNOS) performs the magic
The skin is a solar powered battery!
Cholesterol and Cholesterol Sulfate
Sulfation makes
cholesterol watersoluble and
therefore much
easier to transport
SULFATE
Think about Sulfate!
• Cells in the skin produce vitamin D3 sulfate upon
exposure to the sun
– The precursor to vitamin D3 is cholesterol
• Cells also produce an abundance of
cholesterol sulfate
– I believe this is the more important molecule!
• Many of the alleged benefits of vitamin D3 are
actually benefits of cholesterol sulfate
– Protection against cancer, diabetes and cardiovascular
disease; improved immune function
Heart Disease Mortality and Sunlight*
*Grimes et al., Q. J. Med. 1996; 89:579-589
Vitamin D3 Supplements are
Not the Answer!!*
• 151 people w/ diagnosed
vitamin D deficiency and
heart disease risk profile
• 50,000 IU vitamin D3
supplement for 8 weeks
• Negative results
– Increased serum calcium levels
– Reduced serum parathyroid hormone levels
– Increased serum LDL levels
* P.P. Manish et al., Arteriosclerosis, Thrombosis and Vascular Biology, Epub, 2012
http://www.medpagetoday.com/Cardiology/Dyslipidemia/34561.
Ultraviolet Exposure and Mortality
among Women in Sweden*
• 38,472 Women selected in
1992, aged 30-49
1991-
– monitored for 15 years
• Questionnaire asked about
frequency of sunbathing vacations and sunburn
– Increased sunburn frequency associated with reduced
all-cause mortality
– Sunbathing vacations more than once a year reduced
risk to cardiovascular disease and mortality
* Yang et al., Cancer Epidemiol Biomarkers Prev. 20(4):683-690, 2011
Skin Melanoma Increasing 2%/Yr since 1974
This corresponds to a 30-fold
increase in the use of sunscreen
Andrew Schneider, Aol News, May 24, 2010
aolnews.com/2010/05/24/study-many-sunscreens-may-be-accelerating-cancer
A Biological Definition of Aging
Excess glycation
damage to
proteins (AGEs)
Cholesterol and
sulfate depletion in
cell membranes
CELLS
Dysfunctional blood
system leading to blood
clots and hemorrhages
Increased susceptibility
to infection and cancer
Diabetes and Obesity
Several Roads to Hell
Insufficient
cholesterol sulfate
synthesis in the skin
Steal sulfate from
wherever it’s found
Arthritis, Crohn’s
Disease, colitis,
multiple sclerosis,
type-1 diabetes,
etc.
Synthesize cholesterol
sulfate a different way
obesity,
heart disease
Switch from
cholesterol sulfate to
nitric oxide synthesis
Autism,
Alzheimer’s,
ADHD,
depression
How It’s Supposed to Work
Cholesterol sulfate
synthesized in the skin
upon sunlight exposure
Blood stream delivers
cholesterol and sulfate
to all the tissues
How It Works Instead
Aggressive sun protection
prevents skin from producing
cholesterol sulfate
Cholesterol sulfate is
synthesized in the
atherosclerotic plaque instead
(from homocysteine and LDL)
And/Or …..
From the brain
Alzheimer’s, Multiple Sclerosis
Sulfate is stolen from
whatever tissue is willing
to give it up!
From the gut
(Crohn’s disease, colitis)
From the pancreas
(diabetes, pancreatic cancer)
From the joints (arthritis)
And/Or …..
The body gives up
on sulfate supply
and confronts
blood instabilities
Blood clots
hemorrhaging
Sudden cardiac arrest
Nitric oxide poisoning
Cholesterol Transport
HDL is “good” cholesterol
and LDL is “bad” cholesterol
– true or false?
FALSE!
Neither HDL nor LDL is “cholesterol”!
Both HDL and LDL are particles that
transport cholesterol, fat, fat-soluble
vitamins and antioxidants to all tissues
Some Facts and Speculation
• Fact: Fibroblasts in the skin are major suppliers of
cholesterol to HDL
- Speculation: they depend upon
sulfation for export of cholesterol to HDL
• Fact: Cholesterol sulfate is synthesized
in large quantities in the skin upon sunlight exposure
– Speculation: this is a major supplier of cholesterol and
sulfate to the tissues
– Reduces need for serum LDL
HDL, VLDL, LDL, Chylomicron
• HDL: Liver produces “nascent” particle;
cholesterol-synthesizing cells (fibroblasts) fill it up
o• VLDL: Liver produces large particles; carry fat,
cholesterol, fat-soluble vitamins and antioxidants
o• LDL: “remnant” after VLDL has delivered its goods
• Chylomicron: HUGE particle produced in gut to
deliver ingested fats and cholesterol to the
tissues (mainly heart, liver and muscles)
o
Cholesterol Transport
Bile
Gut
Cholesterol
Synthesis
Fibroblasts
Adrenal Glands
HDL
LDL
Supplies fat and
cholesterol to
the tissues
VLDL
HDL is like a rowboat
that can go in to shore to
pick up goods and
deliver them to the
ocean liner, which is the
VLDL particle
VLDL
HDL
Cholesterol Ester Transport (CEPTP)
Inhibitors*
• This is a new class of drugs that raises
HDL dramatically
HDL
– It does so by trapping cholesterol in the
HDL particle
• These drugs have failed miserably in
trials
CETP
VLDL
– Problem is that they prevent HDL from
doing its job: delivering cholesterol from
the cells in the skin that make it to the
VLDL particles
* Barkowski and Frishman, Cardiol Rev. 2008 16(3):154-62.
Sizes of Lipoprotein Particles in
Blood Serum
Fats and cholesterol enter the
bloodstream from the
digestive system via the
AWESOME
Chylomicron
VERY BAD: small dense LDL
“BAD” “GOOD”
Structure of LDL
Subject to glycation damage
which interferes with
delivery of goods to tissues
Unesterified Cholesterol in
membrane protects
contents from damage and
helps fight infection
Glycation Damage
• Due to excess sugars in the blood stream
– Especially high fructose corn syrup
• Messes up cholesterol transport
– Glycated LDL is less efficient at
delivering cholesterol to tissues
– Glycated LDL can’t be recycled through
receptors in liver (LDLR)
– Builds up in blood serum as
“small dense particles” (The bad kind)
• Cells become deficient in cholesterol
LDL
Cholesterol Sulfate Protects LDL*
sulfate
* E.H. Epstein et al., Science, 214(4521, 659-660, Nov. 6 1981
The Life Cycle of the Chylomicron
Heart,
muscle and
fat cells
• Triglyceride composition closely resembles dietary intake
Dietary Cholesterol
• Dietary fat and cholesterol enter
the lymph system directly from the
gut and exit into the blood stream
at the subclavian vein
• They travel in a large lipid particle
called a chylomicron
• It’s a short path from this major
vein to the heart
• Conclusion: the heart gets first dibs
on dietary cholesterol and fat
Cholesterol Metabolism
adrenal glands
CETP
All the tissues
heart, liver
Cholesterol Transport to HDL
• Cells in the skin are major producers of
cholesterol sulfate and major suppliers of
cholesterol to HDL-A1
• Cells synthesize cholesterol in Endoplasmic
Reticulum (ER)
• Transport from ER to cell membrane
– Is not vesicular
– Ergo, requires migration through cytoplasm
– Proposal: sulfation of cholesterol necessary step in
transport (renders molecule water-soluble)
Cholesterol Management in the Cell*
Acetyl Coenzyme A
LDL
Free cholesterol
ER
HDL
lysosome
ER
Fat stores
Requires sulfation
to cross through
cytoplasm?
Esterified
cholesterol
* E.M.L. Bastiaanse, Cardiovascular Research 33, 272–283, 1997
Sulfate and Cholesterol Transport
• Researchers are puzzled as to how cholesterol is
transported from the ER, where it is synthesized,
to the cell membrane
– Cholesterol’s insolubility in water is problematic
• I propose that the solution is to add sulfate to
make the molecule water-soluble
– This allows cholesterol
(as cholesterol sulfate) to move
freely through the cytoplasm
– This is how synthesized cholesterol
makes it to the cell membrane
AND how it gets from the membrane
to HDL, the particle that transports
it in the blood stream
Sulfate Gives Cholesterol
Free Passage!
HDL
cholesterol
cell
HDL
Low cholesterol
content in HDL is
the strongest lipid
predictor of heart
disease
Cholesterol Sulfate Deficiency
Explains Autistic Correlates
• Autism is associated with several health issues:
– Eczema, asthma, digestive problems (leaky gut)
– Increased susceptibility to infection
• Cholesterol sulfate in skin stimulates filaggrin synthesis
– Filaggrin deficiency leads to eczema and asthma
• Sulfate protects cells in blood from bacteria
• Sulfate deficiency leads to leaky gut syndrome
Nakae et al. The FASEB Journal 22:782.2, 2008
Waring and Klovrza, J Nutr & Environ Medicine 10, 25-32, 2000.
Recent Papers
“Might cholesterol sulfate deficiency contribute to
the development of autistic spectrum disorder?”
Stephanie Seneff, Robert Davidson, Luca Mascitelli
Medical Hypotheses 2012, 78, 213–217
“Impaired Sulfate Metabolism and Epigenetics: Is
There a Link in Autism?”
Samantha Hartzell and Stephanie Seneff
Entropy 2012, 14, 1953-1977
Recapitulation
• HDL, LDL, and chylomicron provide cholesterol
and fat to all the tissues
– Chylomicron supplies dietary cholesterol preferentially
to the heart
• High serum LDL is an indication of insufficient
sulfate and excess glycation damage
• Sulfate is essential for export of cholesterol to
HDL particles
• Cholesterol sulfate deficiency can explain autism
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
Sulfate Deficiency
The body depends upon heparan sulfate
proteoglycans (HSPGs) as a
temporary
storage bin for glucose
Impaired sulfate synthesis in the skin disrupts this
process and leads to diabetes
Heparan Sulfate Proteoglycans
(HSPGs)
Heparan Sulfate: Wonder Worker
Polymers of sugars with attached nitrogen and sulfates: safe glucose storage
A Provocative Hypothesis
When cholesterol and sulfate are in
short supply, storage of sugars in
HSPGs becomes impaired
Cells store glucose temporarily in
HSPGs, protected by sulfate
This is the source of
insulin
resistance and diabetes!
Sulfated Glycosaminoglycans (GAGs)
• Prominent in
extracellular
matrix of all cells
• Amount of
sulfate depends
on availability
• Crucial for
maintaining
negative charge
and protecting
from infection
http://www.science-autism.org/sulphate.htm
Amazing Number of Roles for
Heparan Sulfate in Cells*
* J.R. Bishop et al, Nature 446, 1030-1037, Apr. 2007
Recycling HSPGs w/ LDL*
• Hypothesis:
Sugars are
temporarily
housed in HSPGs
GLUCOSE
LDL
• Hypothesis: sulfate
in HSPGs is
critically needed to
recycle LDL
* J.R. Bishop et al, Nature 446, 1030-1037, Apr. 2007
LDL Uptake by Liver depends on Sulfate in
Heparan Sulfate Proteoglycans (HSPGs)*
• LDL recycled mainly through uptake by liver
• This uptake depends critically on sulfate in HSPGs
• Insufficient HSPGs leads to elevated serum LDL
“Our studies have shown that HSPG participate
not only in the initial sequestration step, but
also in the uptake step, either in association
with the LRP or acting alone as a receptor.”
* Mahli and Ji, J. Lipid Research 40, 1999, 1-16.
LDL Clearance Depends on HSPGs*
LDL Remnant
* Figure 1, in Mahli and Ji, J. Lipid Research 40, 1999, 1-16.
Proposed Role of eNOS in
Heparan Sulfate synthesis
Heparan sulfate
synthesized in
Golgi apparatus
eNOS
cell
Hypothesis: depends upon eNOS
AND SUNLIGHT to add sulfate
Recapitulation
• Heparan sulfate proteoglycans (HSPGs) are
everywhere in the body
– They play a crucial role in ion transport, nutrient
uptake, and cell signalling
– They are central in LDL reuptake and recycling
• I propose that they also provide a temporary
storage depot for glucose
– Deficient sulfate leads to diabetes
• I propose that eNOS provides the sulfate,
catalyzed by sun exposure
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
Obesity
Insufficient HSPGs impairs muscles’ ability
utilize glucose as a fuel
to
Fat cells insinuate themselves into the loop by
transforming glucose to fat for later release to
supply safe usable fuel to muscles
Fat Cells to the Rescue
The U.S. Obesity Epidemic*
1990
2008
2004
WHY ARE WE GETTING
FATTER AND FATTER?
* Source: CDC Behavioral Risk Factor Surveillance System
The Metabolic Syndrome
•
•
•
•
•
Abdominal obesity
Insulin resistance
High blood pressure
High serum triglycerides
High LDL ("bad" cholesterol),
especially “small dense LDL”
• Low HDL ("good" cholesterol)
• Increased risk to heart disease
The Path to Obesity and Heart Disease
Too little cholesterol
sulfate synthesis in
the skin
Sulfate synthesis in artery
wall from homocysteine
(inflammation required)
Platelets synthesize
cholesterol sulfate in
artery wall
Impaired
HSPG
synthesis
Excess
blood
sugar
Glycated LDL
Fat cells take up
damaged LDL and
store cholesterol
Fat cells release
cholesterol to
HDL-A1
Fat cells consume sugar
and store it as fat
Fat cells
proliferate
Fat from fat cells becomes
important fuel for
muscles, including heart
Fat Cells
Get Sick!
Fat Cells Get Sick!
• Cholesterol depletion in
membrane induces cholesterol
synthesis and storage
• Fat cells need sulfate to
release
fat stores
• Too much cholesterol stuck
inside the cell becomes
problematic
• Endoplasmic reticulum has
trouble folding proteins
• Fat cell calls in macrophages to orchestrate cell death
and clean up the mess
Experiment on
Cholesterol Depletion in Fat Cells*
Normal fat cells from rats were exposed in vitro to MβCD
• Depleted cholesterol from cell walls
• Caused activation of cholesterol-acquiring proteins
• Caused insulin resistance and increased synthesis of
TNF-α, angiotensinogen, and IL-6
– Related to hypertension, inflammation, and atherosclerosis
Conclusion:
fat cell membrane cholesterol
depletion is key manifestation
of metabolic syndrome
* Soazig Le Lay et al., J. Biol. Chem., 2001
Endoplasmic Reticulum Becomes Sick
• Endoplasmic reticulum essential for protein
folding and activation
• Absence of sulfate leads to internal
cholesterol accumulation
• Excess cholesterol destroys
cell’s ability to fold proteins
• This causes major
dysfunction and
eventually leads to
cell disintegration
Picture from Wan et Al., The FASEB Journal, 2007
Fat Cells Become Dysfunctional
• Endoplasmic
Reticulum has too
much cholesterol
which inhibits
calcium transport
• Plasma
membrane has
too little
cholesterol,
which causes
potassium leaks
Endoplasmic Reticulum:
Cholesterol Overload
x
Ca+2
Plasma Membrane:
Cholesterol Depleted
K+
Fat Cells Launch Distress Cascade
Macrophage
• Release IL-6 and TNF-α
- Calls in macrophages
- Induces inflammation
F
F
F
F
• Release leptin
- Decreases glucose uptake
• Release angiotensin-II
Distressed Fat Cell
- Suppresses synthesis of apoE
- Increases thirst due to sodium leaks
- Promotes uptake of cholesterol from HDL for
plasma membrane
Visceral Fat
• Damaged VLDL remnants leads to
widespread cholesterol deficiency
• Major organs are especially
susceptible (e.g., pancreas, liver,
adrenal glands, reproductive
organs, etc.)
• Hypothesis: Visceral fat deposits
store reserve cholesterol supply
for critical organs
Putting it All Together
(2)
F
(1)
ER
E
(3)
H
AT-II
F
ER
(5)
(4)
F
F
F
F
“Reverse Epidemiology”*
• Obesity, hypertension, and high cholesterol are
all protective in:
–
–
–
–
–
–
Heart failure
Rheumatoid arthritis
AIDS
Post myocardial infarction
Cancer
The elderly
• This is estimated to apply to 30 million Americans
* George T. Griffing, MD, Medscape Today, Aug 19, 2009.
http://www.medscape.com/viewarticle/707334?src=mp&spon=22
While these measures are risk
factors for heart disease, they
play a protective role in
preventing other consequences
that are arguably worse than
heart disease
17 Year Study on Elderly*
• Begun in 1990: all subjects
were at least 70 years old
• Measured serum cholesterol,
ability to synthesize cholesterol,
and ability to absorb cholesterol
through the intestines
• Low values of all three parameters
were associated with accelerated
mental decline and increased physical frailty
• Subjects with low values on all three had 4 ½
years decreased life span
* Tilvis et al., Annals of Medicine, Early Online, 2011
AIDS Increases Risk to Heart Disease*
“Patients with HIV with chronic infection have
significant vascular inflammation, and thus
added CVD [cardiovascular disease] risk,
beyond that estimated by traditional risk
factors.”
I propose that AIDS is a
cholesterol sulfate deficiency disease
* S. Subramanian, et al., Arterial Inflammation in Patients With
HIV. JAMA, July 25, 201, 308( 4379), p. 386
High Fat Diet Protects from
Heart Failure*
*From W.C. Stanley et al., Dietary Fat and Heart Failure: Moving From Lipotoxicity to
Lipoprotection, Circulation Research 2012, 110:764-776
Recapitulation
• Fat cells play an important service in cleaning up
busted LDL and supplying cholesterol and fats to
insulin-resistant muscle cells
• This is a dirty job, and fat cells get sick
– Too little membrane cholesterol, too much internal
cholesterol
• Macrophages are called in to clean up dying cells
– This leads to inflammatory response and widespread
tissue damage
• Metabolic syndrome is protective in several modern
diseases and conditions
• High fat diet protects from these diseases
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
eNOS
eNOS (endothelial Nitric Oxide Synthase) is a
“moonlighting” enzyme: it makes sulfate upon
sunlight stimulation, and switches to
nitric
oxide (nitrate) under stress
Endothelial nitric oxide synthase
(eNOS)
eNOS!!
• Endothelial nitric oxide synthase (eNOS) is a
very interesting molecule
• It’s known for its role in synthesizing nitric
oxide (NO) from L-arginine
• But I think it has a much more important role
as well, which is its primary role:
– To synthesize sulfate from sulfur in the presence
of sunlight
N  NO  NO3-2
S  SO2  SO4-2
Where is eNOS Found?
What does it do?
• In keratinocytes in the skin
• In the endothelial cells lining
artery walls
• In several cell types in the blood:
– red blood cells, platelets, mast cells
• eNOS synthesizes nitric oxide from L-arginine
It makes no sense for a red blood cell to
synthesize nitric oxide!!
The red blood cell keeps the substrate for
Hemoglobin
to out:
nitric oxide:
nitricbinds
oxidestrongly
synthesis
this
neutralizesthat
the effect
bothto
ofmake
them!
I hypothesize
it usesofeNOS
sulfate instead !
L-ARGININE
eNOS
eNOS and Caveolae
Michel and Feron, J. Clin. Invest. 100(9) 2146-2152, 1997
eNOS with and without L-Arginine
If either L-arginine substrate
or BH4 cofactor is reduced,
eNOS synthesizes O2(superoxide) instead of NO
O2- and NO combine to make
OONO-, a potent oxidizing
agent
Katusic, Am J Physiol Heart Circ
Physiol 281:H981-H986, 2001.
eNOS Dimer Synthesizes Sulfate:
A proposal
• Dimer (two eNOS molecules) creates cavity
• Zinc atom in cavity creates positive charge
field
• Sulfurs in cysteine peptides
electrons
HEME
FLAVINS
attracted to zinc
• Flavins respond to sunlight by
Sulfur Sulfur
emitting electrons
+ +
+Zinc
sulfate
+ ++
Sulfur
• Electrons create charged
Sulfur
oxygen dimers (O2-)
FLAVINS
HEME
electrons
• Oxygen combines with sulfur
to form sulfate (capture energy)
Recapitulation
• eNOS is a moonlighting enzyme whose day job is
cholesterol sulfate synthesis
– Depends on sunlight to catalyze reaction
– Switches to nitric oxide (nitrate) synthesis under
pathological conditions
• Red blood cells, platelets, endothelial cells,
keratinocytes and other cells use eNOS to make
sulfate
– Mechanism involves zinc atom in cavity in eNOS dimer
– These cells need sulfate to produce cholesterol sulfate
and/or heparan sulfate
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
SiNiC
The slide from Sulfur into Nitrogen into Carbon as
we age
Oxygen transport in the blood depends upon these
three atomic elements (S, N, C)
which can
react with oxygen and form anions that stabilize
the blood and safely carry oxygen
Sulfur is the healthiest choice;
nitrogen is
associated with many pathologies; carbon acting
alone will lead to
acidosis and cancer
Signaling Gas Molecules and Oxygen
Sulfur-based
H2S
2O2-
into
Nitrogen-based
NO
H2O2
H2SO4
Superoxide
Superoxideand
or hydrogen
peroxide
are both
highly
peroxide
reacts
reactive
andtosources
with a gas
produceofa
damage
H2inflammation
NO3 stable anion
into
Carbon-based
SiNiC
CO
H2O2
H2CO3
Mancuso et al., J. Neurochem. 113, 563–575, 2010
Signaling Gases
• Hydrogen sulfide (H2S), nitric oxide (NO), and
carbon monoxide (CO) are all released into the
blood stream in small amounts
• All of these gases have the effect of relaxing
the artery wall and increasing oxygen supply
to the tissues
Hydrogen Sulfide as a Signaling Molecule
“It appears that H2S possesses all of the
positive effects of NO without the
capacity to form a toxic metabolite.”
D. Lefer, PNAS 104(46), 17907–17908, 2007
Hydrogen Sulfide Protects from
Cardiovascular Disease*
• BCA blocks H2S
Synthesis
• This causes white
blood cell
adherence to the
artery wall and
infiltration beyond
it, leading to
atherosclerosis
* Renada et al., The Faseb Journal, 20, E1411-E1418, 2006.
eNOS and Caveolae*
When calcium enters the cell,
eNOS stops making sulfate
and starts making nitrate instead
* Michel and Feron, J. Clin. Invest. 100(9) 2146-2152, 1997
Transition to Nitrogen-based
Oxygen Transport*
PI3K
eNOS Requires L-Arginine
VEGF
Interior
as Substrate for NOCell
Synthesis
PI3K
Ca+2
Ca+2
Caveola
Ca+2
CaM
Ca+2 +2
Ca
+2
Ca
Ca+: Calcium
eNOS
NO: Nitric Oxide
Ca+2
CaM: Calmodulin
VEGF: Vascular endothelial growth factor
PI3K: Phosphatidylinositol-3-kinase
eNOS: endothelial nitric oxide synthase
NO
NO
Akt
NO
NO
NO
* Navarro et al., The Faseb Journal 18, 2004
Protamine Treatment*
• Protamine is a cationic
polypeptide highly
Protamine
causesiseNOS
detach
from
Excess
nitric oxide
highlytotoxic,
especially
enriched
in L-arginine
the
membrane
and startconditions
under
well-oxygenated
•profusely
Protamine treatment
can lead to extreme
producing nitric oxide
systemic hypotension,
pulmonary
hypertension,
anaphylactic shock and
cardiac failure
* Viaro et al., Chest 122;1061-1066, 2002
The Relationship with Pathogens
Keep the pathogens out 2O
H2S
2
Fight the pathogens H O
2 2
NO
Let the pathogens winH O
2 2
CO
H2SO4
H2NO3
H2CO3
How it Works
• Keep the pathogens out (sulfur)
– Both cholesterol sulfate in the skin and sulfate
ions surrounding individual cells protect from
bacterial invasion
• Fight the pathogens (nitrogen)
– Once the pathogens get in, nitric oxide is a good
toxin to kill them
• Let the pathogens win (carbon)
– When cells themselves are too vulnerable to
damage, it’s better to just learn to live with the
pathogens –> autoimmune disease
Interleukins Control Strategy
IL-1, TNF-α;
superoxide
IL-6; muscle
loss, anemia
H2S
NO
2O2-
H2O2
IL-10; autoimmunity H O
2 2
CO
H2SO4
H2NO3
H2CO3
How Insufficient Sulfate
Leads to Anemia*
• Bacterial infections rage in blood stream
– Triggers IL-6 synthesis by macrophages
• Induces mechanisms to deplete blood of iron
– Iron feeds the bacteria!
• Iron is drawn into macrophages, to lure
bacteria to their death through nitric oxide
poisoning
• Serum iron is depleted  anemia
Paradkar et al. Blood, 112:3, 866-874, 2008
How Bacterial Infection
Leads to Anemia through IL-6
bacteria
hepcidin
IRON
hepcidin
IRON
IL-6
IRON
IRON
IRON
IRON
Paradkar et al. Blood, 112:3, 866-874, 2008
Conditions Associated
with Deficiencies
Obesity, heart disease, arthritis
-
H2S
2O2
Anorexia, cachexia, ALS
H2O2
NO
Lupus, AIDS, cancer H O
2 2
CO
H2SO4
H2NO3
H2CO3
Recapitulation
• Three signaling gases relate to three systems
of oxygen/acid management: S, N, and C
• Adequate sunlight exposure to skin allows S
system to thrive
• When S is deficient, bacteria invade
• N based system fights bacteria
• C based system gives up 
autoimmune disease
BREAK!
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
Blood Dysfunction
The stability of the blood colloidal system depends
upon adequate cholesterol sulfate.
When sulfate is depleted, suspended cells and
particles become deficient in negative charge, and
start to stick together.
This leads to blood clots and hemorrhaging
Blood Clots and Hemorrhages
Blood Clots and Disease
“How and when our blood clots is one of those
incredibly complex and important processes in
our body that we rarely think about. If your
blood doesn't clot and you cut yourself, you
could bleed to death, if your blood clots too
much, you could be in line for a heart attack
or stroke. “*
* http://medicalxpress.com/news/2012-03-mystery-blood-clotting.html
“Platelet Up-and-Comers”*
“The quest for ever-more effective drugs to
decrease platelet aggregation and reduce the
risk of thrombus formation, without unduly
increasing the risk of bleeding, will always be a
holy grail of cardiovascular disease research.
With each new contender comes fresh
excitement as well as more questions.”
* http://www.theheart.org/collection/Antiplatelet-up-and-comers.do
Hemostasis
• Hemostasis keeps the blood in circulation and
prevents excessive blood loss upon injury
• Maintained by
coagulationinsufficient
system involving clotting
Hypothesis:
factors
sulfate supply to blood leads
• Complex coagulation cascade involving calcium,
to fragile
state of constant
thromboplastin,
phospholipids
and several serum
proteins tension between hemorrhage
– Extrinsic pathwayand blood clots
– Intrinsic pathway
– Common pathway
• Final product is insoluble fibrin (blood clot)
The Glycocalyx
• Negatively charged gel-like mesh lining the walls
of all arteries, veins and capillaries
• Depends crucially on sulfated polysaccharides
– Particularly heparan sulfate proteoglycans (HSPGs)
• Sulfate creates exclusion zones
– Helps protect cells in wall from ion leaks and contact
with enzymes suspended in blood
– Greatly reduces amount of flowing blood (decreases
resistance, lowers blood pressure)
The Glycocalyx
Figure 1 from Weinbaum
et al., Annu. Rev.
Biomed. Eng. 2007.
9:121–67
The Glycocalyx
Sulfate maintains negative charge and creates exclusion zones
Heparan sulfate
proteoglycans
(HSPGs)
Figure 1 from Weinbaum et al., Annu.
Rev. Biomed. Eng. 2007. 9:121–67
Key Role for Sulfates
• Sulfates decorate exterior of cell, e.g., as heparan sulfate
proteoglycans (HSPGs) or as cholesterol sulfate (Ch-S)
• Sulfates carry negative charge and keep cells from sticking
together
• Sulfates, as kosmotropes, create exclusion zone – gel-like
environment to protect cell
Red Blood Cell
Zeta Potential
• Zeta potential indicates degree
of repulsion between similarly
charged particles in a dispersion
(e.g., the blood)
• High zeta potential confers stability
– RBCs and platelets resist aggregation
• Low zeta potential causes flocculation and coagulation
(blood clot)
• Proposal: a steady drop in zeta potential in the blood as
we age is the source of many modern diseases
Grounding the Human Body
Reduces Blood Viscosity*
• Blood viscosity is strongly
influenced by the surface charge
on the suspended particles
(like RBCs)
• A higher repulsive surface charge
increases spacing between RBCs, reduces
clumping, lowers viscosity, and lowers
peripheral resistance to flow (blood pressure)
• Grounding transfers electrons from soil to the
body
– Increases zeta potential, lowers blood viscosity
*G. Chevalier et al., J Alternative and Complementary Medicine, 1–9, 2012
Negative Charge Builds on Artery Wall*
- - - -- - - - -- --- -- -- - - --- - - - -
Red blood cells export cholesterol sulfate to the
artery wall, supplying it with cholesterol, sulfate,
and negative charge
* Davidson and Seneff, Entropy 14, 1399-1442, 2012.
A Battery!
* Davidson and Seneff, Entropy 14, 1399-1442, 2012.
Battery Poles Between Artery and Vein
“current”
ARTERY
VEIN
• RBC’s lose charge as they travel through the capillary
• This sets up voltage gradient between vein and artery
• Negatively charged RBC’s are propelled towards positive
pole of “battery”
This creates force field that promotes blood flow
* Davidson and Seneff, Entropy 14, 1399-1442, 2012.
Zeta potential measures rate at which
negatively charged particles travel in an
electric force field
Hypothesis
• When blood becomes deficient in negative
charge (sulfates), glycocalyx becomes
unhealthy
• Endothelial cells develop gaps that allow
blood to seep into tissues
• Blood clots are needed to plug the holes
• Mistakes can lead to dangerous blockage
– Deep vein thrombosis
– Pulmonary thrombosis
Recapitulation
• Hemostasis is the regulatory system that maintains
blood stability
– Tension between clotting and hemhorraging
– Sulfates in the glycocalyx are protective
• Grounding can improve blood stability by
providing negative charge through currents from
the ground
• Red blood cells discharge negative charge by
releasing cholesterol sulfate in capillaries
– This helps maintain pH difference between arteries and
veins to drive circulation
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
Heart Disease
Cardiovascular plaque develops as an alternative
mechanism to produce cholesterol sulfate from
damaged LDL and homocysteine
What Happens when Cholesterol
Sulfate Synthesis is Impaired??
Activities in Plaque Produce
Cholesterol Sulfate to Supply the Heart
They Knew a Long Time Ago*
• Article published in 1960
• Fed cholesterol to monkeys
– induced atherosclerosis
• If sulfur-containing nutrients
are added, atherosclerosis is
prevented
• These nutrients provide
source of sulfate to enable
cholesterol transport
* G.V. Mann et al., Am. J. Clin. Nutr. 8, 491-497, 1960.
Steps in Atherosclerosis*
1. Inflamed endothelium provides adhesion
molecules to trap and hold macrophages
2. Macrophages through scavenger process take
up oxidized LDL and become foam cells
3. Interleukins and growth factors promote
proliferation of smooth muscle cells
(artery thickening)
4. Extracellular matrix proteins are degraded
5. Vulnerable plaque eruption: thrombosis
* Libby et al., Circulation 105:1135-1143, 2002
Steps in Atherosclerosis
Entrapment of
macrophages
Build up of
fatty deposits
Rupture and
thrombosis
Adapted from Libby, et al., Circulation 105:1135-1143, 2002
Many Good Reasons for ROS
• ROS (reactive oxygen species) are a key
component of inflammation in the artery
• ROS are needed to produce sulfate
• Oxidation of glycated LDL makes it accessible
to macrophages for breakdown
• Peroxynitrite (product of reaction between
superoxide and nitric oxide) is toxic to
pathogens
Mitsuhashi et al. Shock 24(6) 529-34, 2005.
Kaplan and Aviram, Arterioscler Thromb Vasc Biol 21(3) 386-93 2001.
Alvarez et al., J. Biol. Chem. 286, 6627-6640, 2011.
Macrophages and Cholesterol*
• Macrophages in artery wall take up oxidized LDL
and export extracted cholesterol to HDL-A1
• Unsaturated fatty acids interfere with export
process
• Macrophages eventually
become damaged by
exposure to oxidizing
and glycating agents 
necrotic core
* Wang and Oram, J. Biol. Chem. 277 (7) , 5692–5697, 2002
Quote from the Abstract*
“These findings raise the possibility that an increased supply of unsaturated fatty acids in the
artery wall promotes atherogenesis by impairing
the ABCA1 cholesterol secretory pathway in
macrophages.”
* Wang and Oram, J. Biol. Chem. 277 (7) , 5692–5697, 2002
Pathway from Homocysteine to Sulfate*
Vitamin A
homocysteine
thiolactone
Superoxide:
Reactive oxygen species
(ROS)
alpha keto
butyrate
When folate and
B12 are deficient,
nearly all
homocysteine is
converted to
homocysteine
thiolactone
thioretinamide
Vitamin C
sulfite
retinoic acid
Sulfite
oxidase
PAPS
sulfate
*McCully, Annals of Clinical and
Laboratory Science 41:4, 300-313, 2011
Synthesis of PAPS consumes ATP
eNOS’ synthesis of sulfate upon
sunlight exposure in skin may avoid
loss of ATP by exploiting energy
from light instead
Platelets and Cholesterol Sulfate
• Platelets and RBCs both synthesize
cholesterol sulfate (Ch-S)
– Ch-S is present in the
atherosclerotic lesions in the aorta
– Platelets will accept cholesterol only from HDL-A1
– Platelet synthesis rate increases 300-fold when PAPS is
available.
– PAPS is formed from ATP and sulfate
• Platelet aggregation leads to thrombosis
– HDL suppresses aggregation; LDL promotes it
Yanai et al, Circulation 109, 92-96, 2004
Putting it All Together
homocysteine
Heart
muscle
cell
sulfate
Synthesize sulfate
ATP
PAPS
Red
Blood
Cell
Lipid
Raft
Cholesterol
protects
the
heart
from
RBC
ion leaks and sulfate
allows it
insulin
Platelet
to
safely metabolizeglucose
glucose
(3)
macrophage
Ch
Artery Wall
Ch
ApoE
ROS
Inflammatory Agents
Endothelial Cell
Steps in Atherosclerosis:
Reinterpreted
1. Endothelial cells lining artery walls feeding
the heart release inflammatory agents
2. Macrophages infiltrate artery wall
3. Macrophages extract cholesterol from
oxidized LDL and deliver it to HDL-A1.
4. Platelets extract cholesterol from HLD-A1 and
convert it to cholesterol sulfate, with help
from PAPS
5. Macrophages die and build up necrotic core
I hypothesize that treatments aimed at
reducing the supply of cholesterol to the
plaque will eventually lead to severe
deficiencies in cholesterol and sulfate supply
to the heart, resulting in heart failure.
National Heart, Lung, and Blood Institute National Institutes of Health Data Fact Sheet
Recapitulation
• I believe that cardiovascular disease can be
best characterized as a factory to supply
cholesterol and sulfate to the heart
• Unsaturated fats, but not saturated fats,
interfere with the supply chain
• Statin drugs, through their ability to deplete
the supply of cholesterol to the plaque, can
lead to heart failure down the road
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
Impaired Gut Bacteria
Gut Bacteria and Health*
• We are an ecosystem: a community of
interacting cells
• Trillions of bacteria live in our body
• They outnumber our own cells 10:1
• Killing them off may make us sick or fat
• 5-8 million microbial genes in our bodies
(3 million in digestive system from more
than 1000 species)
• They play very essential roles in our bodies
– Digest foods, absorb nutrients, provide
enzymes, make vitamins and
antiinflammatories
– Regulate appetite and brain function
• Vaginal birth matters
• Everyone has unique set of gut bacteria
* NPR’s On Point: Your Inner Ecosystem, http://onpoint.wbur.org/2012/06/20/bacteria-2
“Our microbes are under threat —
and the enemy is us”*
“Microbiome”
• Microbes in our body outnumber us 10:1
– 99% of them are benign or offer protection
– Synthesize vitamins B12 and folate
• Birth through birth canal (v.s. C section)
– Infant picks up mother's microbiome
• Colostrum provides food for gut bacteria
• Industrialized countries are too clean
– Drastically altered microbiome
• H. pylori 90% --> < 10% in last hundred years in U.S.
• Esophageal cancer and childhood asthma have risen in step
* Liz Szabo, USA Today,
http://www.usatoday.com/news/health/story/2012-07-13/body-bugs-microbes/56255904/1
H. pylori: There are Benefits!*
• The Western world has now significantly reduced
the presence of H. pylori in the gut
– This has reduced the incidence of stomach ulcers and
stomach cancer
– But it has also increased the incidence of esophageal
reflux disease and esophageal cancer
• H. pylori serve a useful role in reducing acid
production by stomach: excess acid aggravates
esophagus
• We host a colony of bacteria and they work
together with us for a jointly beneficial solution
*Caroline Hadley, EMBO reports 7(5), 471-473, 2006
“Folate synthesized by bacteria in the
human upper small intestine is
assimilated by the host”*
• Folate synthesized by flora in the small intestine is
absorbed through the gut
• Atrophic gastritis associated with higher duodenal pH
– Leads to bacterial overgrowth
– Increased supply of folate
• This could be an important
benefit to assure adequate
folate
* E. Camilo et al., Gastroenterology 110(4), 991-998, 1996
Wild Speculation
• Leaky gut and leaky skin and
leaky blood brain barrier lead
to infiltration of microbes
from gut or skin into brain
• Bacteria are “lured” into the
body & into the brain and
then killed and harvested to
renew important nutrients
• Possible nutrients include:
– Vitamin B12 (cobalamin), vitamin B9 (folate),
thiamine, vitamin K and heparan sulfate
“The Infection Connection”*
• Barry Marshall drank H. pylori broth and
developed gastritis
– Proved that H. Pylori “causes” stomach ulcers
• Other examples:
– Cervical cancer & HPV
– Liver cancer & Hepatitis B and C
– Lymphoma & Epstein-Barr virus
– Cardiovascular disease & chronic infection?
– Alzheimer’s & brain infection?
*Caroline Hadley, EMBO reports 7(5), 471-473, 2006
What Causes Increased Infection?
Impaired Cholesterol Sulfate Synthesis!
Microbes
Microbes
Microbes
Leaky Gut
• Collitis and Crohn’s disease are associated
with significantly reduced sulfate in the GAGs
in the walls of the intestines
• Cells are defective in transporting nutrients
– Leads to severe vitamin deficiencies (e.g., B12)
• Tight junctions between cells become loose
• Microbes can penetrate the gut wall and enter
the blood stream
– They can be harvested by macrophages!
Microvilli in Gall Bladder Epithelium*
Healthy – negative charge makes them stand tall
Unhealthy – addition of cationic polymer destroys them
* Figures 1 and 2 in Quinton and Philpott, J. Cell Biol. 56, 1978, 787-796.
Bacteria can Swim Across Gut Barrier!
Microbial Invasion through BBB*
* Figure 5, p. 380, J.A. Orellana et al., Antioxidants and Redox Signaling 11(2), 2009
The Good Side of Infection and
Inflammation in the Brain
• Inflammation activates microglia to
phagocytose dying cells and Amyloid beta
• Bacterial exposure promotes phagocytosis of
dying cells
• Both dying cells and bacteria can be recycled
into new raw materials!
– These nutrients may be essential to brain health
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
Infection
Susceptibility to infection is a consequence of
impaired cholesterol sulfate synthesis, which
introduces widespread pathology
Infection serves a useful role in
resupplying
critical nutrients such as
cobalamin,
folate, and heparan sulfate
The Falling Apart Syndrome*
• The key message: All chronic
diseases can be linked to infections
• The author, Russell Farris,
previously published another
book:
– The Potbelly Syndrome: How
Common Germs Cause Obesity,
Diabetes and Heart Disease.
* Russell Farris, http://www.polymicrobial.com
Figure from Chapter 6 in
The Potbelly Syndrome*
* By Russell Farris
A Possible Cause-and-Effect Relationship
• First comes impaired barriers, impaired
nutrient transport and impaired blood
colloidal suspension
• Certain nutrients are desperately needed to
fix the problems: e.g., cobalamin, vitamin K,
heparan sulfate
– Bacteria are invited in to provide those nutrients!
• The disease may be uncomfortable, but the
alternative may be worse!
Infection and Atherosclerosis*
* A. Tufano et al., Semin Thromb Hemost 2012(38), 515–523.
What Happens if you Treat
with Antibiotics?*
* A. Tufano et al., Seminars in Thrombosis & HemostasisVol. 38(5), 2012, 515-523.
What Happens if you Treat
with Antibiotics?*
Significantly more people die of
cardivascular disease following
antibiotic treatment (p=0.01)
* A. Tufano et al., Seminars in Thrombosis & HemostasisVol. 38(5), 2012, 515-523.
Chlamydia Produce Heparan Sulfate!*
• Chlamydia are viable only
inside host cells
• They set up housekeeping
in vacuoles within the cell
(e.g., a macrophage in the plaque)
• They produce a glucosamine-containing
sulphated polysaccharide that is nearly
indistinguishable from heparan sulfate
* S.J. Rasmussen-Lathrop et al, Cell Microbiol. 2000 Apr, 2(2), 137-44.
Special Issue Devoted to Alzheimer's
and Infection:* Key Points
• Pathogens can produce progressive chronic diseases
like Alzheimer’s, asthma, and heart disease
• Alzheimer himself proposed involvement of infective
agents in Alzheimer's 100 years ago
• Pathogens stimulate inflammation
• Pathogens evade host defenses and establish chronic
latent infections
• Persistent superoxide, nitric oxide and peroxynitrite
(ROS) cause DNA damage and apoptosis and alter gene
expression
• Environmental toxins and poor nutrition weaken
immune system and provide opportunity to bacteria
and viruses
* Special Issue of the Journal of Alzheimer’s Disease, 2008
Amyloid Plaque Structure*
• Microglia and amyloid plaque
accumulate side-by-side in a
central region surrounded by
astrocytes
• Microglia harbor dormant
bacteria
• Should the bacteria leave,
they will encounter the
plaque, which will kill them
• The astrocytes guard the gates
and shield the neurons from
the damaging plaque
Chlamydia in plaque region
produce heparan sulfate!
* Microphotograph from Schwab et
al., Journal of Alzheimer’s Disease 13
359–369, 2008.
Amyloid Beta and Nitric Oxide*
• Aβ produced by the neuron induced
increased synthesis of nitric oxide
• Nitric oxide reacts with superoxide
to produce peroxynitrite
• Peroxynitrite is extremely damaging
to mitochondria and disrupts their
function
• Cell commits apoptosis due to
insufficient energy supply
* U. Keil et al., J. Biol. Chem, 2004 279(48), 50310-50320
Sepsis and the Vasculature*
Impaired ability to
modulate blood flow
Immune cells stick
to wall of venule
and block flow
Small blood clots
form in capillaries
and block flow
* S. Trzeciak et al., Acad Emerg Med. 2008 May ; 15(5): 399–413
Hypothesis
Blood flow needs to nearly shut down
during sepsis; organs need to shut down;
the entire body focuses on harvesting
heparan sulfate from the invasive
microbes to recover blood stability.
GSK-3: Inducing Inflammation*
• GSK-3 is a protein that interferes with insulinmediated glucose uptake and with glucose
storage into glycogen
• It also promotes inflammation in association
with mood disorders, Alzheimer’s disease,
diabetes, and cancer
• This shows a clear link between glucose
impairment and inflammation
* R.S. Jope et al., Neurochem Res. 2007 ; 32(4-5): 577–595.
Figure 2 from R.S. Jope et al., Neurochem Res. 2007 ; 32(4-5): 577–595.
Inflammation and
impaired glucose
metabolism are
closely linked
Inflammation is
needed to generate
more sulfate!
Figure 2 from R.S. Jope et al., Neurochem Res. 2007 ; 32(4-5): 577–595.
Recapitulation
• Infection is associated with many chronic diseases like
heart disease and Alzheimer’s
– Treating with antibiotics doesn’t help
– Infection serves useful role in supplying nutrients?
– Support comes from Chlamydia’s ability to produce
heparan sulfate
– Amyloid beta plaque provides home for microbes
– Sepsis reflects blood melt-down due to sulfate deficiency
• Inflammation and impaired glucose metabolism are
closely linked
– Linking element is sulfate deficiency?
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
Impaired Autophagy
Autophagy is the process by which cells dispose
of the garbage that accumulates with living – they
can recycle misfolded proteins into new proteins
and replace broken mitochondria with fresh ones.
Excess nitric oxide leads to impaired autophagy,
which results in accumulation of debris and busted
mitochondria. Over time, the cell becomes so
impaired that it has to shut down.
Inflammation and Repair*
• Inflammation is at the root of a host of diseases,
from diabetes to cancer
– Induces permeability of vascular walls and movement
of immune cells into the tissues
– Leads to production of ROS and tissue damage
• Acute inflammation follows infection or tissue
injury
• Chronic inflammation characterizes diseases like
heart disease, diabetes, and Alzheimer's disease
• Cell debris is phagocytized by leukocytes
* Kolattukudy, and Niu, Circulation Research 2012, 110:174-189
MCP-1 is a Key Signaling Molecule*
“The increased level of serum MCP-1 found in
humans correlated with markers of the metabolic
syndrome, including obesity, insulin resistance,
type 2 diabetes, hypertension, and increased
serum concentration of triacylglycerol.”
* p. 181, Kolattukudy, and Niu, Circulation Research 2012, 110:174-189
Inflammation and ER Stress*
MCP-1
Cell Death
Progenitor
Cells Build
New Tissues
* Figure 3, Kolattukudy, and Niu, Circulation Research 2012, 110:174-189
A Picture Emerges …
Insufficient cholesterol sulfate synthesis in the skin
Long-term
recovery
Inflammatory agents
(TNF-α, LPS, IL-1)
MCP-1
Emergency
reaction
NFκB
MCPIP
NO, ONOOHeart disease
obesity
Cholesterol sulfate restored
Heparan sulfate restored
Mitochondrial dysfunction
Autophagy dysfunction
Unfolded protein response
Autophagy
• Allows cell to recycle damaged goods
• Both damaged mitochondria and damaged
endoplasmic reticulum are absorbed into the
lysosome and broken down
• Autophagy depends on sulfates in HSPGs
http://www.wormbook.org/chapters/www_autophagy/autophagy.html
The Lysosome is Central to the Cell*
* M. Jeyakumar, et al., Nature Reviews 6, Sep. 2005, Box 1, p. 2
Recycling HSPGs w/ LDL*
• Hypothesis:
Sugars are
temporarily
housed in HSPGs
GLUCOSE
LDL
• Hypothesis: sulfate
in HSPGs is
critically needed to
recycle LDL
* J.R. Bishop et al, Nature 446, 1030-1037, Apr. 2007
Fasting Induces Autophagy
• Upregulation of autophagy is
neuroprotective
• Much effort has been
expended trying to develop
drugs that pass through the
blood brain barrier and
enhance autophagy in the
brain
• Short-term fasting leads to a
dramatic upregulation in
neuronal autophagy
M. Alirezaei et al., Autophagy 6(6):702-710, Aug 16, 2010.
Fasting Induces Autophagy
• Upregulation of autophagy is
neuroprotective
• Much effort has been
“Our
data to
lead
us to speculate that
expended
trying
develop
sporadic
fasting might
drugs that
pass through
the represent a
blood brain
barrier
andinexpensive means
simple,
safe and
enhance autophagy
in this
the potentially
to promote
brain
therapeutic neuronal response”
• Short-term fasting leads to a
dramatic upregulation in
neuronal autophagy
M. Alirezaei et al., Autophagy 6(6):702-710, Aug 16, 2010.
Autophagy and Insulin
• Autophagy is suppressed by insulin
– Insulin resistance leads
to impaired autophagy
• Impaired autophagy may
explain build-up of
amyloid beta and tau
protein in senile plaque
of Alzheimer’s disease
• Impaired autophagy is also associated with
metabolic syndrome, cancer, asthma, and cardiac
hypertrophy
H.-Y. Liu et al., JBC 284(45):31484–31492,2009
Nitric Oxide and Autophagy
• Autophagy is a natural mechanism by which
cells clean up debris and dysfunctional
organelles
• Excess nitric oxide inhibits autophagy
• Switchover to nitric oxide by eNOS leads to
autophagy dysfunction and accumulation of
garbage in the cell
• Long-term autophagy inhibition leads to
cellular dysfunction and cell death
Some Effects of Excess Nitric Oxide
on the Brain*
•
•
•
•
Impairs autophagy
Damages lipids in cell membranes
Disrupts mitochondrial synthesis of ATP
Interferes with reuptake of serotonin,
dopamine, and epinephrine
*O. Akyol et al., In Vivo 18: 377-390, 2004
Common Underlying Pathology*
NITRIC OXIDE
CALCIUM
Applies to autism, Alzheimer’s disease, Parkinson’s disease,
ALS, depression and seizures,
as well as lysosomal storage disorders (rare genetic diseases)
* Jeyakumar et al., Nature Reviews:Neuroscience 6, Sep. 2005.
Recapitulation
• Autophagy is the process by which cells clean up their
accumulating debris
– Insulin and nitric oxide suppress autophagy
– Fasting enhances it
• Autophagy and phagocytosis are closely related in that
both take place in the lysosome, and the lysosome
depends on both cholesterol and sulfate to function
properly
• Autophagy failure is a common pathology associated
with many diseases
– Leads to broad restructuring of process of cell
maintenance and explains the onset of these diseases
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
Environmental Toxins
Some Environmental Toxins
•
•
•
•
•
•
PCB’s (plastic)
Pesticides
Perfluorooctanoic Acid (PFOA) (teflon)
Glyphosate (Roundup)  hydroxybenzoic acid
Arsenic
Aluminum
Synergy between Environmental Toxins
and Epigenetics*
• Epigenetics is interface between inherited genome
and environment
• Epigenetic dysregulation is increasingly implicated
in disease
– Dutch Hunger Winter (1944-45) associated with
hypomethylation of IGF2 leading to diabetes in
offspring several decades later
– Hypomethylation also associated w/ arthritis, heart
disease autism, and cancer
• We are exposed to a growing number of
environmental toxins
– Particularly lipophilic xenobiotics like PCB’s, dioxins,
pesticides
* D.-H. Lee et al., Environmental Health Perspectives 117(12) Dec. 2009, 1799-1802.
Synergy between Environmental
Toxins and Epigenetics* (Cont’d)
• Dietary deficiencies compound effects
– S-adenosyl methionine (SAM) (methyl donor) requires
methionine, folate, choline, betaine, and B12
– Mother’s deficiencies may induce epigenetic bias
• Chronic low-grade exposure to environmental
toxins depletes glutathione
– Methionine is precursor to glutathione 
methionine drain  methylation impairment 
cancer, etc.
* D.-H. Lee et al., Environmental Health Perspectives 117(12) Dec. 2009, 1799-1802.
Impaired Sulfate Metabolism and
Epigenetics: Is There a Link in Autism?*
DNA
Hypomethylation
methylation
Methionine
Methionine
Sulfate
• Methionine sits at the crossroads of the
pathway to sulfate and to DNA methylation
• Decreased sulfate synthesis in the skin leads
to depletion of methionine (drain to sulfate)
• This induces hypomethylation of DNA in utero
and reprogramming towards a model of a
sunless world  autism
* S. Hartzell and S. Seneff, , Entropy, 14, 1953-1977.
“Perfluorooctanoic Acid and
Cardiovascular Disease in US Adults”*
• PFOA is a toxin and carcinogen found in Teflon and Gore-Tex
• Increasing serum levels of PFOA are associated with
increased risk to cardiovascular disease
* A. Shankar et al., Archives of Internal Medicine, 2012.
sciencedaily.com/releases/2012/09/120903221128.htm
Glyphosate
• Glyphosate is now the #1 herbicide in the U.S.
and is increasingly used around the world
– Developed and patented by Monsanto in the 1970’s
– Came out from patent in 2000
– Inhibits an enzyme involved in synthesis of tyrosine,
tryptophan and phenlalanine (the three aromatic
amino acids)
• Surfactant POEA (polyethoxylated tallow amine)
is often included in products, and is highly toxic
to animals and people
Benzene Toxicity!
• Glyphosate disrupts synthesis of aromatic
amino acids
• Leads to build-up of intermediate product:
shikimate  4-hydroxybenzoic acid
• This is a toxic compound!
• Glyphosate is claimed to be relatively safe
because humans don’t have shikimate
pathway
• However, gut bacteria do!!!!
Sulfate-reducing Bacteria can Detoxify
Benzene Compounds*
• Study involved anaerobic bacteria in marine
sediment
• Analogy can be drawn with human colon
– Sulfate-reducing bacteria are overabundant in association
with autism
– Are they needed to detoxify benzene?
Benzene + sulfate + water  carbonate + hydrogen sulfide
* F. Musat and F. Widdel, Environmental Microbiology 10(1), 10–19, 2008.
Arsenic
• Arsenic exposure leads to
increased risk to cardiovascular
disease*
• Soil bacteria chelate arsenic
through sulfate synthesis**
• My hypothesis: glyphosate
(Roundup) kills soil bacteria and
permits arsenic accumulation
* Y..C. Hsieh et al., Environ Res. 2011 Aug, 111(6), 804-10.
** M.F. Kirk et al. Geology; Nov. 2004, 32(11), 953-956
Quote From Abstract*
“Where sulfate-reducing bacteria are active,
the sulfide produced reacts to precipitate
arsenic, or coprecipitate it with iron, leaving
little in solution. In the absence of sulfate
reduction, methanogenesis is the dominant
type of microbial metabolism, and arsenic
accumulates to high levels.”
* M.F. Kirk et al. Geology; Nov. 2004, 32(11), 953-956
Consumer Reports (CR) Study on Rice*
76% of US Rice
Grown in Cotton Belt
* http://www.consumerreports.org/cro/arsenicinfood.htm
Summary of CR Study
• Samples from American Cotton
Belt had higher levels than
those from other countries
• Brown rice higher than white
rice
• People who ate rice had 44%
greater body burden of arsenic
• Action: Asking Food and Drug
Administration to set limits in
rice products and fruit juices
Aluminum: The Worst Toxin?
We are Too Comfortable
with Aluminum!
"Aluminum is not perceived, I believe, by the
public as a dangerous metal and, therefore,
we are in a much more comfortable wicket in
terms of defending its presence in vaccines”
-- Dr. John Clement, World Health
Organization, San Juan, Puerto Rico,
May 11th - 12th, 2000, p. 64
Aluminum and Alzheimer’s Disease
• Aluminum is a neurotoxin that inhibits more than
200 biologically important functions
• Aluminum has been associated with multiple
diseases affecting the nervous system:
–
–
–
–
Dialysis encephalopathy
ALS (Lou Gehrig's disease)
Parkinsonism dementia in Guam
Alzheimer's disease
• Aluminum may play crucial role as a cross-linker
in amyloid beta oligomerization
Kawahara and Kato-Negishi, International J. Alzheimer's Disease, Article ID 276393,2011
Aluminum, Dialysis, and Dementia*
• Patients with end-stage
kidney disease
• Aluminum in water
in the dialysate
• Leads to severe dementia
• Occurs after three to five
years of dialysis
• Can be avoided by careful filtering of water supply
* Wills and Savory, Environmental Health Perspectives 63, 141-147, 1985
Aluminum’s Many Effects in the Brain
Kawahara and Kato-Negishi, International J. Alzheimer’s Disease 2011, Article ID 276393
Aluminum Exposure and
Memory, Depression*
• 25 symptomatic workers from the same
aluminum smelting plant
– 22 (88%) reported frequent loss of balance
– 21 (84%) reported memory loss
– 21 (84%) showed physical signs of incoordination
• 19 were tested for depression on the
Minnesota Multiphasic Personality Inventory
– 17 (89%) tested positive for depression
*White et al., Arch Intern Med. 152(7):1443-1448, 1992.
Our Studies with VAERS Database on
Aluminum & Depression
• VAERS: Vaccine Adverse Event Reporting
System, maintained by CDC
• Tabulate word frequencies for mentions of
"depression" in adverse reactions to
aluminum-containing vaccines versus
non-aluminum-containing vaccines.
– 231 mentions in aluminum-containing vaccines
versus 85 in age-matched controls
– Highly significant result (p = 0.005)
Aluminum’s effect on Red Blood Cells*
• Aluminum causes microcytic anemia
– Induces severe morphological changes in RBCs
– Leads to eryptosis – cell dies
• N-acetyl cysteine (a source of sulfur) affords
some protection
Normal
With Aluminum
* D.M. Vota et al., Journal of Cellular Biochemistry 113:1581–1589, 2012.
Antiperspirants and Breast Cancer *
A: antiperspirant
D: deodorant
S: shaving
* K.G. McGrath, European Journal of Cancer Prevention 2003, 12:479–485
Antiperspirants and Breast Cancer *
A: antiperspirant
“Ninety per
cent of the United States
D: deodorant
populationS: shaving
regularly uses antiperspirants
and deodorants, with their daily use
($ sales) only exceeded by toothpastes”
* K.G. McGrath, European Journal of Cancer Prevention 2003, 12:479–485
Aluminum Foil in Cooking*
• Leaching from aluminum foil into foods during
cooking may carry significant health risks
– Provides an easy channel for metal
entry into the body
• Factors leading to more leaching:
– Higher temperature
– Higher acidity
– Salt and spices
• Particularly problematic for people with
leaky gut syndrome
* G. Bassioni, Int. J. Electrochem. Sci., 7 (2012) 4498 - 4509
Aluminum in Vaccines:
In 1999,
industry Gamble*
A vaccine
Neurological
responded to autism parents’
concerns about mercury, but
simultaneously began increasing
aluminum exposure
* Neil Z. Miller, Thinktwice Global Vaccine Institute, www.thinktwice.com
“Empirical Data Confirm Autism Symptoms Related
to Aluminum and Acetaminophen Exposure”*
• Hypothesis: Vaccine industry removed
mercury from vaccines but simultaneously
augmented aluminum burden
• This is what prevented autism rates from
falling following mercury reduction
• MMR association with autism is in part
attributable to administering Tylenol to curb
fever
– Tylenol detoxification depletes sulfate
* S. Seneff, R.M. Davidson, and J. Liu, Entropy, Special Issue on Semiotics, 2012
Recapitulation
• We live with multiple environmental toxins
• Two that I am particularly worried about are
aluminum and glyphosate
• Aluminum likely plays a role in autism,
Alzheimer’s disease, depression and breast
cancer
• We have become complacent with these
toxins, and they are slowly eroding our
collective health
Outline
• Introduction
– Big Ideas: Dysfunction and Consequences
• Dysfunction
–
–
–
–
–
Cholesterol Transport
Sulfate Deficiency
Obesity
Endothelial Nitric Oxide Synthase (eNOS)
SiNiC
• Consequences
–
–
–
–
–
Blood Clots and Hemorrhages
Cardiovascular Disease
Impaired Gut Bacteria
Infection
Impaired Autophagy
• The Environment
– Environmental Toxins
– Polyphenols
• Summary
Polyphenols
Polyphenols
•
•
•
•
•
Tannin in tea
Flavonoids in fruits
Resveratrol in wine
Curcumin in curries
Polyphenols in coffee and chocolate
These compounds all have
significant health benefits, but it’s
not clear why!
Polyphenols in Coffee*
"Supplementation with CPP [Coffee
Polyphenols] significantly reduced body
weight gain, abdominal and liver fat
accumulation, and infiltration of
macrophages into adipose tissues."
* T. Murase et al., Am J Physiol Endocrinol Metab 300: E122–E133, 2011
Four Related Molecules
Curcumin
Resveratrol
All of the OH groups
can become sulfates
Benzene
Phenol
“High Absorption but Very Low
Bioavailability of Oral Resveratrol in Humans”*
• Researchers studied absorption of resveratrol
• Wanted to understand its role in biology
• Found that it was sulfated in the gut, went
into the bloodstream and promptly
disappeared
• I hypothesize that its health benefit has solely
to do with its ability to transport sulfate
*T. Walle et al., Drug Metabolism and Disposition, 32(12), 1377-1382, 2004
Vitamin C Sulfate!
OH
OH
Recapitulation
• A variety of “healthy” compounds in foods
have the interesting property that they can be
sulfated
• These include coffee, tea, resveratrol (wine),
flavonoids in berries and other fruits,
curcumin in curries and vitamin C
• I maintain that the health benefits of these
foods are solely due to their ability to
transport sulfate in the blood stream
Summary
Modern Environment
Depletes Sulfate Everywhere
Sunscreen blocks
sunlight and disables
eNOS function
Modern Environment
Depletes Sulfate Everywhere
Glyphosate induces toxic
phenols that deplete
sulfate in their breakdown
by colon bacteria
Modern Environment
Depletes Sulfate Everywhere
Depleted sulfur in our food
supply due to modern
farming practices
Modern Environment
Depletes Sulfate Everywhere
Vaccines deplete sulfate via
aluminum and mercury
detoxification; these toxins
also interfere with eNOS
function
Modern Environment
Depletes Sulfate Everywhere
Various pharmaceutical
drugs like acetaminophen
deplete sulfate in their
detoxification process
Modern Environment
Depletes Sulfate Everywhere
Antibiotics kill gut bacteria
that we depend on for critical
bioactive molecules like
cobalamin and vitamin K
What You Can Do!
Eat Foods Containing Sulfur!
Go Organic!
Probiotics!
Escape to a sunny place in winter!
If you Live in Canada, Use a Sunlamp!
Epsom Salts!
Magnesium Sulfate in hot bath
water is a cheap and easy way to
get sulfate supply to the skin
Don’ts!
Summary
• I have identified impaired cholesterol sulfate
synthesis as a key factor in the diseases of
modern times
– I have shown how this can lead to diabetes, obesity,
autism, Alzheimer’s disease and heart disease
• Microbes are often unsung heroes who help
rescue us from pathological situations of our own
making
• Environmental toxins play an important role in
our current state of health crisis
Thank you!